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A Blanchard

Publications and source records attributed to A Blanchard.

86 records · Page 5Linked to original sources
PubMed1990

[Infectivity inhibition of HIV prototype strains by antibodies directed against a peptide sequence of mycoplasma].

Antibodies prepared against a peptide corresponding to the site of cyto-adherence of Mycoplasma genitalium adhesine inhibit or reduce the infectivity of the HIV-1BRU and HIV-2ROD strains of Human Immunodeficiency Virus in lymphoid cells. These results strengthen the hypothesis that some mycoplasmas may play an important part in HIV replication and pathogenicity.

Amino Acid Sequence↗
PubMed1989

Topology and acylation of spiralin.

Of the 51 polypeptides detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the plasma membrane of the helical mollicute Spiroplasma melliferum, 21 are acylated, predominantly with myristic (14:0) and palmitic (16:0) chains. This is notably the case for spiralin, the major membrane protein of this bacterium, which contains an average of 0.7 acyl chains per polypeptide, attached very probably by ester bonds to alcohol amino acids. The amphiphilicity of spiralin was demonstrated by the behavior of the protein in charge-shift electrophoresis, its incorporation into liposomes, and its ability to form in the absence of lipids and detergents, globular protein micelles (diameter, approximately 15 nm). The presence of epitopes on the two faces of the cell membrane, as probed by antibody adsorption and crossed immunoelectrophoresis, and the strong interaction between spiralin and the intracytoplasmic fibrils show that spiralin is a transmembrane protein. The mean hydropathy of the amino acid composition of spiralin (-0.30) is on the hydrophilic side of the scale. Surprisingly, the water-insoluble core of spiralin micelles, which is the putative membrane anchor, has a still more hydrophilic amino acid composition (mean hydropathy, -0.70) and is enriched in glycine and serine residues. Taking into account all these properties, we propose a topological model for spiralin featuring a transbilayer localization with hydrophilic domains protruding on the two faces of the membrane and connected by a small domain embedded within the apolar region of the lipid bilayer. In this model, the membrane anchoring of the protein is strengthened by a covalently bound acyl chain.

Acylation↗
PubMed1988

Ubiquitin is a common factor in intermediate filament inclusion bodies of diverse type in man, including those of Parkinson's disease, Pick's disease, and Alzheimer's disease, as well as Rosenthal fibres in cerebellar astrocytomas, cytoplasmic bodies in muscle, and mallory bodies in alcoholic liver disease.

Polyclonal antibodies were raised which have a high affinity for conjugated ubiquitin. Immunocytochemistry was performed on paraffin sections of tissues showing well-characterized inclusion bodies. Ubiquitin was found as a component of the intermediate filament inclusion bodies characteristic of several major diseases including Lewy bodies of Parkinson's disease, Pick bodies of Pick's disease, Mallory bodies of alcoholic liver disease, cytoplasmic bodies of a specific myopathy, and Rosenthal fibres within astrocytes. Ubiquitin was also present in the three histological lesions characteristic of Alzheimer's disease. These observations suggest a fundamental role for ubiquitin in the formation of intermediate filament inclusion bodies in man, and have implications regarding the pathogenesis of these important diseases.

Alzheimer Disease↗
PubMed1988

Characteristics of Ureaplasma urealyticum urease.

Sonication of Ureaplasma urealyticum cells grown in a dialysate growth medium effectively separated the cytoplasmic fraction from the membrane fraction, with both fractions relatively free from exogenous contaminating proteins. The urease activity was associated with the cytoplasmic fraction, and the ureaplasmal urease exhibited a specific activity higher than that of crystalline jack bean urease. The enzymatic activity of the ureaplasmal enzyme was optimum at pH 7.5 and was resistant to the chelating agents EDTA and sodium citrate. Sulfhydryl-blocking agents such as HgCl2 and Pb(NO3)2 inhibited the ureaplasmal urease, which was also shown to be particularly sensitive to flurofamide and, to a much lesser extent, to acetohydroxamic acid. Electrophoretic analysis of the proteins of the ureaplasmal cell fractions combined with Western immunoblot with an antiserum to the ureaplasmal urease indicated that the urease constitutes a major component of the cytoplasm and is composed of several 70-kilodalton polypeptides.

Citrates↗
PubMed1987

Localization of spiralin in Escherichia coli cells transformed with the recombinant plasmid pESI.

The expression of spiralin in the transformant strain HB101 Tsp of Escherichia coli has been investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), immunodetection after electrotransfer, and crossed immunoelectrophoresis. The protein has been sought in whole cells, cytoplasm, and plasma and outer membranes of the bacterium. Untransformed E. coli cells and Spiroplasma citri cells were used as negative and positive references, respectively. Contrary to earlier claims, spiralin was detected not only in the cytoplasm of E. coli, but also in the inner and outer membranes. In addition, our results show that the protein was not secreted by the bacterium. Three forms of spiralin could be distinguished with respect to differences in apparent molecular mass: 27.5, 29.5 and 30 kilodalton (kDa). The presence of the high molecular mass polypeptide in the two membranes of E. coli invalidates the hypothesis according to which the 27.5-kDa (or 28-kDa) species is a mature form derived from the 30-kDa (or 30.5-kDa) species by the amputation of a signal sequence. Since spiralin is an acyl protein, the possibility of variation in the extent of acylation of the protein in E. coli, with subsequent variation of the apparent molecular mass, should be taken into account.

Acylation↗
PubMed1987

Amphiphilic properties of spiralin, the major surface antigen of spiroplasmas. A preliminary report.

Spiralin, a 28-kDa (kilodalton) polypeptide, is the major antigen of Spiroplasma citri and S. melliferum in which it usually represents more than 20% of total membrane protein. The amino acid compositions of the spiralins purified from both spiroplasma species unambiguously show that these proteins are homologous. In addition, several lines of evidence indicate that such a protein is present in the menbrane of S. apis. A 25-kDa polypeptide antigenically related to S. citri spiralin has also been purified from the membrane of the nonhelical variant ASP-1. The spiralin of S. melliferum B88 has been used as a model for extensive characterization. This antigen binds detergent under nondenaturing conditions, can be incorporated into liposomes, and forms protein micelles upon gentle removal of detergent. Digestion of the micelles with trypsin leads to the precipitation of an insoluble material containing a major polypeptide of 3.9 kDa. The amino acid composition of this fragment is different from that of intact spiralin. It is highly enriched in glycine and serine and, as an insoluble peptide, exhibits an unexpectedly high polarity index (PI = 51.4%). Screening for acyl proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodetection in the membrane of S. melliferum indicates that spiralin is actually acylated. This set of properties is evidence that spiralin is an intrinsic membrane protein and strongly suggests that acylation triggers or facilitates integration of the molecule into the lipid bilayer of the spiroplasma membrane.

Acylation↗
PubMed1986

[Ulcerative colitis caused by Histoplasma capsulatum in a patient with the acquired immunodeficiency syndrome].

The authors report a case of ulcerative colitis due to Histoplasma capsulatum in a previously healthy 35 year-old french geologist with acquired immune deficiency syndrome (AIDS). Gomori-Grocott and PAS stains and indirect immunofluorescence revealed Histoplasma capsulatum in colonic biopsies. The search for LAV antibody was positive. T-lymphocyte analysis revealed 10/mm3 OKT4 with OKT4/OKT8 ratio of 0.16. Histoplasmosis should be considered in subjects with ulcerative colitis according to the epidemiological context. In patients with AIDS relapses after discontinuation of treatment are to be expected and suppressive therapy with an imidazole derivative should probably be continued indefinitely.

Acquired Immunodeficiency Syndrome↗
PubMed1985

Highly selective extraction of spiralin from the Spiroplasma citri cell membrane with alkyl-N-sulfobetaines.

The extraction of proteins from the membrane of the mollicute (mycoplasma) Spiroplasma citri by sodium N-dodecyl-N,N-dimethyl-3-amino-1-propane sulfonate (SB12) and sodium N-tetradecyl-N,N-dimethyl-3-amino-1-propane sulfonate (SB14) was studied with electrophoretic methods. The membranes were prepared by osmotic lysis of the cells and depleted of the bulk of extrinsic proteins. It was possible to extract up to 35 and 45% of membrane proteins with SB12 and SB14, respectively. Maximal yield was obtained in both cases with detergent concentrations greater than or equal to 5 mumoles/mg of membrane protein. Spiralin, the major protein in the S. citri membrane, was highly selectively solubilized without the loss of antigenicity, with a yield of about 90% with SB12 and close to 100% with SB14, for a detergent concentration greater than or equal to 0.2 M. The degree of selectivity in favour of spiralin was higher with SB12 (purity approximately equal to 70%) than with SB14 (purity approximately equal to 50%). Treatment of the S. citri membrane with high concentrations of SB12 is a simple and fast procedure for partial purification of spiralin. This example shows that, in some cases, it should be possible to modulate the selectivity of the extraction of membrane proteins simply by varying the relative concentration of detergent.

Bacterial Outer Membrane Proteins↗
PubMedDate not supplied

Cloning of Ureaplasma urealyticum DNA sequences showing genetic homology with urease genes from gram-negative bacteria.

DNA chromosomic sequences from Ureaplasma urealyticum were shown, by DNA hybridization, to possess homology with Providencia stuartii urease genes. The homologous ureaplasma DNA fragment was cloned in Escherichia coli and a DNA probe, designated IC61 probe was specific for only the urease-producing U. urealyticum among the Mollicutes. Genetic homology shown between the IC61 probe and urease genes from Campylobacter pylori, E. coli and P. stuartii suggests that (1) the cloned DNA fragment from U. urealyticum contains urease encoding sequences, and that (2) the urease genes possess conserved sequences among phylogenetically distant prokaryotes.

Cloning, Molecular↗
PubMedDate not supplied

Comparison of the amino acid compositions and antigenic properties of spiralins purified from the plasma membranes of different spiroplasmas.

Spiralins were purified by agarose-suspension electrophoresis after extraction with detergents from the membranes of the following spiroplasmas: Spiroplasma citri C189, S. citri Maroc (R8A2), S. citri Scaph and the honey-bee spiroplasma B88. The four proteins (molecular mass congruent to 26,000 daltons, as determined by sodium dodecyl sulphate-pore gradient electrophoresis) showed very similar amino acid compositions characterized by the absence of methionine and tryptophan and a high polarity index (greater than 49%). When compared with the amino acid composition of S. citri membrane, the four spiralins had little or no histidine, a low content of glycine, leucine, tyrosine, phenylalanine and arginine, and a high content of threonine, alanine and valine. Comparison of the amino acid compositions according to the criteria described by Cornish-Bowden (Anal. Biochem., 1980, 105, 233-238) strongly suggests that all four spiralins are related. A crossed immunoelectrophoretical comparison, however, shows that though the three proteins purified from S. citri strains (serogroup I-1) are antigenically similar, they do not seem to share common epitopes with spiralin from the honey-bee spiroplasma B88 (serogroup I-2).

Amino Acids↗